Targeting IKK beta and aurora kinases in melanoma

描述(由申请人提供)： 目前黑色素瘤的流行主要是由于年龄、阳光和化学暴露而影响我们退伍军人中的老年男性和女性。在黑色素瘤的进展过程中，极光激酶A和B的扩增和/或NF-：B的失控导致非整倍体，促进细胞增殖，逃避细胞凋亡，增加炎性介质的产生和肿瘤进展。我们有初步数据表明，当极光A激酶被阻断时，一些人类黑色素瘤肿瘤的异种移植瘤的生长受到抑制，而其他黑色素瘤病变的异种移植瘤对IKK2抑制剂有反应。我们假设IKK2的结构性激活和/或极光激酶(AIK)的放大可以促进黑色素瘤的进展，并且抑制这些途径将抑制肿瘤的生长。此外，我们假设人类黑色素瘤基因表达谱可以预测哪些肿瘤对AIK抑制剂有反应，哪些肿瘤对IKK2抑制剂有反应。本项目有三个具体目标：1)利用组织芯片免疫组织化学分析方法，研究极光激酶A、B、C在黑色素瘤进展过程中的表达特点，并确定IKK/AIK激酶的过表达如何影响黑色素瘤细胞和肿瘤生长；2)确定抑制IKK和AIK通路是否可被证明比单独针对单个激酶更有效地治疗黑色素瘤，并确定抑制IKK和AIK如何影响抗肿瘤宿主防御；3)研究对AIK或IKK2抑制剂起反应或不起作用的人黑色素瘤肿瘤的基因表达谱。利用组织芯片的免疫组织化学分析，我们将研究极光蛋白激酶和磷酸化(relA/p65)在肿瘤进展过程中痣、发育不良痣、原发和转移性黑色素瘤病变的表达模式。我们将利用可诱导表达系统在体外改变AURA和IKK2在黑素细胞和黑色素瘤细胞中的表达，以确定这两条途径过度激活的后果。我们将在小鼠异种移植模型中检测黑色素瘤对IKK2、AIKs及其组合的抑制剂的反应。最后，使用基因表达微阵列分析，我们将确定区分药物应答者和非应答者的表达谱。从这项提案中得出的数据将为更好地设计黑色素瘤患者的临床试验提供关键的见解，这将对我们的退伍军人群体产生积极的影响。 公共卫生相关性： 黑色素瘤是美国生长最快的肿瘤类型之一。黑色素瘤的“流行病”很大程度上仅限于老年男性。我们的退伍军人人口由于年龄、阳光和化学暴露而受到越来越多的影响。我们想了解调节细胞生长和逃脱死亡的阻断激酶[极光激酶和/或kappa-β激酶抑制因子(ikk？)]是如何在黑色素瘤进展过程中表达的，它们如何在肿瘤生长、细胞分裂过程和细胞死亡中发挥作用。我们还将在临床前研究中评估这些激酶及其抑制剂对正常细胞的影响。我们想要描述对极光激酶抑制剂和IKK抑制剂或这两种抑制剂的组合有反应或无反应的黑色素瘤患者的基因表达谱。这项研究的结果将为更好地设计未来的临床试验提供关键信息，这些试验有望表明对患有恶性黑色素瘤或其他癌症类型的退伍军人的治疗有所改善。

DESCRIPTION (provided by applicant): The current melanoma "epidemic" is primarily affecting older men and women in our Veteran population due to age, sun and chemical exposure. During melanoma tumor progression amplification of aurora kinases A and B and/or the deregulation of NF-:B lead to aneuploidy, enhanced cell proliferation, escape from apoptosis, enhanced production of inflammatory mediators and tumor progression. We have preliminary data showing that xenografts of some human melanoma tumors are growth inhibited when aurora A kinase is blocked, while xenografts from other melanoma lesions respond to an IKK2 inhibitor. We hypothesize that constitutive activation of IKK2 and/or aurora kinase (AIK) amplification can contribute to melanoma tumor progression and that inhibition of these pathways will inhibit tumor growth. Moreover, we hypothesize that human melanoma gene expression profiles can predict which tumors will respond to AIK inhibitors and which will respond to IKK2 inhibitors. There are three specific aims for this project: 1) to characterize the expression of aurora kinase A, B, C during melanoma tumor progression using immunohistochemical analysis of tissue microarrays and to determine how the over-expression of IKK/ AIK kinases affect melanocytes and tumor growth; 2) to determine whether inhibiting both IKK and AIK pathways may prove to be more efficacious for treatment of melanoma tumors than by targeting each individual kinase alone and to determine how inhibition of IKK and AIK affect the anti-tumor host defense; 3) to characterize the gene expression profiles for human melanoma tumors that respond or do not respond to AIK or IKK2 inhibitors. Using immunohistochemical analyses of tissue microarrays we will examine the expression pattern of aurora kinases and phosphorylated (RelA/p65) of nevi, dysplastic nevi, primary and metastatic melanoma lesions during tumor progression. We will utilize an inducible expression system to alter the expression of AurA and IKK2 in melanocytes and melanoma cells in vitro to determine the consequences of over-activation of these two pathways. We will examine the response of melanoma tumors to inhibitors of IKK2, AIKs, and combinations thereof in a murine xenograft model. Finally, using gene expression microarray analysis, we will determine the expression profiles that distinguish between drug responders versus non-responders. The data derived from this proposal will provide key insights for better design of clinical trials for melanoma patients which should positively impact our Veteran population. PUBLIC HEALTH RELEVANCE: Melanoma is one of the fastest growing tumor types in the US. The melanoma "epidemic" is largely confined to older men. Our Veteran population is increasingly affected due to age, sun and chemical exposure. We would like to learn how blocking kinases that regulate cell growth and escape from death [aurora kinases and/or inhibitor of kappa-beta kinase (IKK¿)] are expressed during melanoma progression, how they function in tumor growth, cell division processes, and cell death. We also will assess the effects of these kinases and their inhibitors on normal cells during preclinical studies. We would like to characterize the gene expression profile for melanoma patients that respond or do not respond to aurora kinase inhibitors and IKK¿ inhibitors, or a combination of these two inhibitors. Results from this study will provide key information for better design of future clinical trials which will hopefully indicate improved treatment for Veterans experiencing malignant melanoma-or other cancer types.